Contract Award for Topic 388:Development of DNA Aptamer-Based Lateral Flow Assays for Liver Flukes (Phase I)

Nanohmics计划利用市购的华支睾吸虫和乌verrini Opisthorchis抗原的重组表面蛋白和n端肽，开发敏感、特异的DNA适配体胶体金夹层横向流动（LF）试纸条。将通过在中国和泰国已建立的接触者获得实际的肝吸虫进行检测。如果n端序列肽在第一阶段不能被证明是足够的靶标，那么Nanohmics将在第二阶段针对c端肽和测序的吸虫蛋白的富含脯氨酸的弯曲区域开发适配体，这些区域可能标记可达的细胞外结构域。同样的适体可以附着在磁珠上，用于从粪便浆中浓缩吸虫卵或蠕虫，然后进行寄生虫裂解或蛋白质提取和LF检测。如有必要，将采用高荧光量子点报告适体来提高视觉检测灵敏度。在第二阶段，Nanohmics还将开发自己的智能手机控制的LF测试条读取器和软件应用程序，用于在资源有限的环境下通过手机网络报告数据。

Nanohmics proposes to develop sensitive and specific DNA aptamer-based colloidal gold sandwich lateral flow (LF) test strips from commercially available recombinant surface proteins and N-terminal peptides of Clonorchis sinensis and Opisthorchis viverrini antigens. Actual liver flukes for testing will be obtained through established contacts in China and Thailand. If the N-terminal sequence peptides do not prove to be adequate targets in Phase 1, then Nanohmics will develop aptamers in Phase 2 against C-terminal peptides and proline-rich bend regions of the sequenced trematode proteins which are likely to mark accessible extracellular domains. The same aptamers can be attached to magnetic beads and used to concentrate trematode eggs or worms from fecal slurries followed by parasite lysis or protein extraction and LF detection. And if necessary, highly fluorescent quantum dot-reporter aptamers will be employed to enhance visual detection sensitivity. In Phase 2, Nanohmics will also develop its own smart phone-controlled LF test strip reader and software applications for data reporting via the cell phone network in resource-limited environments.